反相高效液相色谱法测定大鼠血浆中左旋黄皮酰胺及其主要代谢产物和药代动力学

目的　探讨左旋黄皮酰胺［（-）clau］及其主要代谢产物6-OH-clau在大鼠体内的药代动力学。方法　建立了RP-HPLC-UV法。固定相为Kromasil-100 C₁₈（5 μm）色谱柱,流动相为乙腈-甲醇-水（21∶16.5∶62.5), DM-9384作内标，氯仿作提取溶剂。结果　测得（-）clau的回收率为96.91%～105.74%,日内、日间RSD低于7%,最低检测浓度为24 ng.mL^-1,（-）clau和6-OH-clau分别在0.047～968 μg.mL^-1和0.049～200 μg.mL^-1,线性关系良好(γ=0.999); （-）clau和6-OH-clau血浆浓度-时间曲线符合二室开放模型,同时求得两者的药代动力学参数。结论　数据表明（-）clau在大鼠血浆中的分布、代谢转化和消除均较快。

DETERMINATION OF PLASMA CONCENTRATION OF (-) CLAUSENAMIDE AND 6-HYDROXYL-CLAUSENAMIDE IN RATS BY HPLC AND THEIR PHARMACOKINETICS

AIM　To Determine the concentration of (-) clausenamide［(-)clau］ and its essential metabolite─6-Hydroxyl- clausenamide(6-OH-clau) in rat plasma and study their pharmacokinetics.METHODS　A reversed-phase high performance liquid chromatography with ultraviolet detection has been developed for determination of (-)clau and its metabolites in rat plasma using a column of Kromasil C₁₈(5 μm) and mobile phase of acetonitrile-methanol-water(21∶16.5∶62.5). DM-9384 was used as an internal standard. Rat plasma samples were extracted with chloroform. RESULTS　The mean relative recovery of (-)clau was 100.9%±4.4%(n=3) with within-day precision of 1.8%～6.2% and between-day precision of 1.06%～3.31%, and the limit of detection was 24 ng.mL^-1. At the range of 0.47～968 μg.mL^-1 for (-)clau, and 0.049～200 μg.mL^-1 for 6-OH-clau, there was a linear relationship with a value for γ=0.999. By the nonlinear least-squares regression and F test between different compartments using a computer program 3P87, the plasma concentration vs time course of (-) clau given iv 80 mg.kg^-1 to rats was found to fit a 2-compartments open model. The following pharmacokinetic parameters were obtained: for (-)clau, T_1/2α=(12.19±1.68) min, Vd=(1.19±0.31)　L.kg^-1, T_1/2β=(156.65±18.65)　min and AUC0～720=(1286.09±171.56)　min.μg.L^-1; for 6-OH(-)clau, T_1/2Kt=(9.61±3.56)　min,　T_1/2β=(238.81±48.66)　min, Tmax=(17.91±3.05)　min, Cmax=(1.29±0.11) μg*mL^-1 and AUC_0～480=(78.31±5.35)　min.μg.L^-1. CONCLUSION　(-)Clau exhibited a pharmacokinetic character that its distribution, transformation and elimination in rat plasma are all relatively rapid.